Detection of ochratoxin A in porcine kidneys by a monoclonal antibody-based radioimmunoassay
| Autor: | Guido Slegers, W. H. Stimson, A. A. G. Candlish, John E. Smith, D. M. Rousseau, C. Van Peteghem |
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| Rok vydání: | 1987 |
| Předmět: |
Ochratoxin A
medicine.drug_class Swine Radioimmunoassay Enzyme-Linked Immunosorbent Assay Monoclonal antibody medicine.disease_cause Kidney Applied Microbiology and Biotechnology Immunoglobulin G Cell Line chemistry.chemical_compound Tissue culture Mice medicine Animals Bovine serum albumin Hybridomas Ecology biology Toxin food and beverages Antibodies Monoclonal Molecular biology Ochratoxins chemistry Cell culture biology.protein Food Science Biotechnology Research Article |
| Zdroj: | Applied and environmental microbiology. 53(3) |
| ISSN: | 0099-2240 |
| Popis: | By using an indirect enzyme-linked immunosorbent assay, eight monoclonal antibodies (MAbs) were selected. Mice were immunized with ochratoxin A that was conjugated to bovine serum albumin. The hybridoma cell line designated 10G2 was grown in tissue culture and as an ascites tumor. The MAb was characterized to be specific to ochratoxin A and of the immunoglobulin G (IgG) class. Subsequently, the ascites fluid of this hybridoma was used in a competitive solid-phase IgG radioimmunoassay on protein A-Sepharose CL-4B, with [14C]ochratoxin A as tracer. Porcine kidneys were extracted with 0.5% phosphoric acid in chloroform. A two-step cleanup was achieved on a Sep-Pak C18 cartridge and a Sep-Pak silica cartridge. Radioimmunoassay with MAbs coupled to protein A-Sepharose CL-4B allowed the detection of ochratoxin A in porcine kidneys at a concentration as low as 0.2 ng/g. |
| Databáze: | OpenAIRE |
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